Evaluation of Two Methods of Isolation, Differentiation and Staining of Lymphocytes by Microlymphocytotoxicity Assay

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Title Evaluation of Two Methods of Isolation, Differentiation and Staining of Lymphocytes by Microlymphocytotoxicity Assay
Avaliação de Dois Métodos de Separação, Diferenciação e Coloração de Linfócitos por meio da Reação de Microlinfocitotoxicidade
 
Creator Rosa, Glicélia da
Oliveira, Alessandra Valéria de
Borelli, Sueli Donizete
Pereira, Luciene Christina Marques da Silva
 
Subject Ciências Biológicas
microlinfocitotoxicidade; prova cruzada; anticorpos; microlymphocytotoxicity; cross match; antibodies
 
Description Microlymphocytotoxicity assay is routinely used to develop the cross match. Donor lymphocytes, in this assay, are target to anti-HLA alloantibodies that could be present in the receptor sera and because of that, is important to define the methodology of isolation, differentiation and staining of lymphocytes. Due to cellular fragility of lymphocytes, mainly B lymphocytes, methodology of isolation, differentiation and staining show direct influence in the reaction. The aim of this study was to compare two methodologies of isolation, differentiation and staining of lymphocytes by ficoll-hypaque/fluorobeads and tripan blue/fluoroquent respectively. Twenty-seven reactions were developed using ficoll-hypaque/tripan and 42 by fluorobeads/fluoroquent methods. The results showed that fluorobeads/fluoroquent method had better performance in the process of isolation, differentiation and staining for both lymphocytes populations, T and B.
A reação de microlinfocitotoxicidade é rotineiramente utilizada para realizar a prova cruzada. Os linfócitos do doador, nesta reação, são alvos dos aloanticorpos anti-HLA que podem estar presentes no soro do receptor, e por isso é importante definir a metodologia de separação, diferenciação e coloração dos linfócitos. Devido à fragilidade celular dos linfócitos, principalmente dos linfócitos B, os métodos de separação, diferenciação e coloração apresentam influência direta no resultado da reação. O objetivo deste estudo foi comparar duas metodologias de separação, diferenciação e coloração de linfócitos: os métodos de ficoll-hypaque/fluorobeads e azul de tripan/fluoroquent, respectivamente. Vinte e sete provas cruzadas foram desenvolvidas utilizando o método de ficoll-hypaque/tripan e 42 provas cruzadas pelo método de fluorobeads/fluoroquent. Os resultados mostraram a superioridade do método fluorobeads/fluoroquent no processo de separação, diferenciação e coloração para as duas populações de linfócitos, T e B.
 
Publisher Centro Universitário de Maringá
 
Contributor
 
Date 2007-11-20
 
Type info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
Avaliado por Pares

 
Format application/pdf
 
Identifier http://periodicos.unicesumar.edu.br/index.php/iccesumar/article/view/546
 
Source Iniciação Científica Cesumar; v. 9, n. 2 (2007); 89-94
2176-9192
1518-1243
 
Language por
 
Relation http://periodicos.unicesumar.edu.br/index.php/iccesumar/article/view/546/463
 

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